Method of using soluble epoxide hydrolase inhibitors

ABSTRACT

Disclosed are novel compounds of the formula Ia and methods of using soluble epoxide hydrolase (sEH) inhibitors of the formulas I and Ia for diseases related to cardiovascular disease.

RELATED APPLICATION DATA

[0001] This application claims benefit to U.S. provisional applicationNo. 60/302,066 filed Jun. 29, 2001.

FIELD OF THE INVENTION

[0002] This invention is directed to methods of using soluble epoxidehydrolase (sEH) inhibitors for diseases related to cardiovasculardisease.

BACKGROUND OF THE INVENTION

[0003] Epoxide hydrolases are a group of enzymes ubiquitous in nature,detected in species ranging from plants to mammals. These enzymes arefunctionally related in that they all catalyze the addition of water toan epoxide, resulting in a diol. Epoxide hydrolases are importantmetabolizing enzymes in living systems. Epoxides are reactive speciesand once formed are capable of undergoing nucleophilic addition.Epoxides are frequently found as intermediates in the metabolic pathwayof xenobiotics. Thus in the process of metabolism of xenobiotics,reactive species are formed which are capable of undergoing addition tobiological nucleophiles. Epoxide hydrolases are therefore importantenzymes for the detoxification of epoxides by conversion to theircorresponding, non-reactive diols.

[0004] In mammals, several types of epoxide hydrolases have beencharacterized including soluble epoxide hydrolase (sEH), also referredto as cytosolic epoxide hydrolase, cholesterol epoxide hydrolase, LTA₄hydrolase, hepoxilin hydrolase, and microsomal epoxide hydrolase(Fretland and Omiecinski, Chemico-Biological Interactions, 129:41-59(2000)). Epoxide hydrolases have been found in all tissues examined invertebrates including heart, kidney and liver (Vogel, et al., Eur J.Biochemistry, 126:425-431 (1982); Schladt et al., Biochem. Pharmacol.,35:3309-3316 (1986)). Epoxide hydrolases have also been detected inhuman blood components including lymphocytes (e.g. T-lymphocytes),monocytes, erythrocytes, platelets and plasma. In the blood, most of thesEH detected was present in lymphocytes (Seidegard et al., CancerResearch, 44:3654-3660 (1984)).

[0005] The epoxide hydrolases differ in their specificity towardsepoxide substrates. For example, sEH is selective for aliphatic epoxidessuch as epoxide fatty acids while microsomal epoxide hydrolase (mEH) ismore selective for cyclic and arene oxides. The primary knownphysiological substrates of sEH are four regioisomeric cis epoxides ofarachidonic acid known as epoxyeicosatrienoic acids or EETs. These are5,6-, 8,9-, 11,12-, and 14,15-epoxyeicosatrienoic acid. Also known to besubstrates are epoxides of linoleic acid known as leukotoxin orisoleukotoxin. Both the EETs and the leukotoxins are generated bymembers of the cytochrome P450 monooxygenase family (Capdevila, et al.,J. Lipid Res., 41:163-181 (2000)).

[0006] The various EETs appear to function as chemical mediators thatmay act in both autocrine and paracrine roles. EETs appear to be able tofunction as endothelial derived hyperpolarizing factor (EDHF) in variousvascular beds due to their ability to cause hyperpolarization of themembranes of vascular smooth muscle cells with resultant vasodilation(Weintraub, et al., Circ. Res., 81:258-267 (1997)). EDHF is synthesizedfrom arachidonic acid by various cytochrome P450 enzymes in endothelialcells proximal to vascular smooth muscle (Quilley, et al., Brit. Pharm.,54:1059 (1997)); Quilley and McGiff, TIPS, 21:121-124 (2000)); Flemingand Busse, Nephrol. Dial. Transplant, 13:2721-2723 (1998)). In thevascular smooth muscle cells EETs provoke signaling pathways which leadto activation of BK_(ca2+) channels (big Ca²⁺ activated potassiumchannels) and inhibition of L-type Ca²⁺ channels. This results inhyperpolarization of membrane potential, inhibition of Ca²⁺ influx andrelaxation (Li et al., Circ. Res., 85:349-356 (1999)). Endotheliumdependent vasodilation has been shown to be impaired in different formsof experimental hypertension as well as in human hypertension (Lind, etal., Blood Pressure, 9:4-15 (2000)). Impaired endothelium dependentvasorelaxation is also a characteristic feature of the syndrome known asendothelial dysfunction (Goligorsky, et. al., Hypertension, 37[part2]:744-748 ( 2001). Endothelial dysfunction plays a significant role ina large number of pathological conditions including type 1 and type 2diabetes, insulin resistance syndrome, hypertension, atherosclerosis,coronary artery disease, angina, ischemia, ischemic stroke, Raynaud'sdisease and renal disease. Hence, it is likely that enhancement of EETsconcentration would have a beneficial therapeutic effect in patientswhere endothelial dysfunction plays a causative role. Other effects ofEETs that may influence hypertension involve effects on kidney function.Levels of various EETs and their hydrolysis products, the DHETs,increase significantly both in the kidneys of spontaneously hypertensiverats (SHR) (Yu, et al., Circ. Res. 87:992-998 (2000)) and in womensuffering from pregnancy induced hypertension (Catella, et al., Proc.Natl. Acad. Sci. U.S.A., 87:5893-5897 (1990)). In the spontaneouslyhypertensive rat model, both cytochrome P450 and sEH activities werefound to increase (Yu et al., Molecular Pharmacology, 2000, 57,1011-1020). Addition of a known sEH inhibitor was shown to decrease theblood pressure to normal levels. Finally, male soluble epoxide hydrolasenull mice exhibited a phenotype characterized by lower blood pressurethan their wild-type counterparts (Sinal, et al., J.Biol.Chem.,275:40504-40510 (2000)).

[0007] EETs, especially 11,12-EET, also have been shown to exhibitanti-inflammatory properties (Node, et al., Science, 285:1276-1279(1999); Campbell, TIPS, 21:125-127 (2000); Zeldin and Liao, TIPS,21:127-128 (2000)). Node, et al. have demonstrated 11,12-EET decreasesexpression of cytokine induced endothelial cell adhesion molecules,especially VCAM-1. They further showed that EETs prevent leukocyteadhesion to the vascular wall and that the mechanism responsibleinvolves inhibition of NF-κB and IκB kinase. Vascular inflammation playsa role in endothelial dysfunction (Kessler, et al., Circulation,99:1878-1884 (1999)). Hence, the ability of EETs to inhibit the NF-κBpathway should also help ameliorate this condition.

[0008] In addition to the physiological effect of some substrates of sEH(EETs, mentioned above), some diols, i.e. DHETs, produced by sEH mayhave potent biological effects. For example, sEH metabolism of epoxidesproduced from linoleic acid (leukotoxin and isoleukotoxin) producesleukotoxin and isoleukotoxin diols (Greene, et al., Arch. Biochem.Biophys. 376(2): 420-432 (2000)). These diols were shown to be toxic tocultured rat alveolar epithelial cells, increasing intracellular calciumlevels, increasing intercellular junction permeability and promotingloss of epithelial integrity (Moghaddam et al., Nature Medicine,3:562-566 (1997)). Therefore these diols could contribute to theetiology of diseases such as adult respiratory distress syndrome wherelung leukotoxin levels have been shown to be elevated (Ishizaki, et al.,Pulm. Pharm.& Therap., 12:145-155 (1999)). Hammock, et al. havedisclosed the treatment of inflammatory diseases, in particular adultrespiratory distress syndrome and other acute inflammatory conditionsmediated by lipid metabolites, by the administration of inhibitors ofepoxide hydrolase (WO 98/06261; U.S. Pat. No. 5,955,496).

[0009] A number of classes of sEH inhibitors have been identified. Amongthese are chalcone oxide derivatives (Miyamoto, et al. Arch. Biochem.Biophys., 254:203-213 (1987)) and various trans-3-phenylglycidols(Dietze, et al., Biochem. Pharm. 42:1163-1175 (1991); Dietze, et al.,Comp.Biochem. Physiol. B, 104:309-314 (1993)).

[0010] More recently, Hammock et al. have disclosed certain biologicallystable inhibitors of sEH for the treatment of inflammatory diseases, foruse in affinity separations of epoxide hydrolases and in agriculturalapplications (U.S. Pat. No. 6,150,415). The Hammock '415 patent alsogenerally describes that the disclosed pharmacophores can be used todeliver a reactive functionality to the catalytic site, e.g., alkylatingagents or Michael acceptors, and that these reactive functionalities canbe used to deliver fluorescent or affinity labels to the enzyme activesite for enzyme detection (col. 4, line 66 to col. 5, line 5). Certainurea and carbamate inhibitors of sEH have also been described in theliterature (Morisseau et al., Proc. Natl. Acad. Sci., 96:8849-8854(1999); Argiriadi et al., J. Biol. Chem., 275 (20) 15265-15270 (2000);Nakagawa et al. Bioorg. Med. Chem., 8:2663-2673 (2000)).

[0011] WO 99/62885 (A1) discloses 1-(4-aminophenyl)pyrazoles havinganti-inflammatory activity resulting from their ability to inhibit IL-2production in T-lymphocytes, it does not however, disclose or suggestcompounds therein being effective inhibitors of sEH. WO 00/23060discloses a method of treating immunological disorders mediated byT-lymphocytes by administration of an inhibitor of sEH. Several1-(4-aminophenyl)pyrazoles are given as examples of inhibitors of sEH.

[0012] As outlined in the discussion above, inhibitors of sEH are usefultherefore, in the treatment of cardiovascular diseases such asendothelial dysfunction either by preventing the degradation of sEHsubstrates that have beneficial effects or by preventing the formationof metabolites that have adverse effects. Further investigation by thepresent inventors has shown that the inhibition of IL-2 production andinhibition of sEH are separable activities with divergentstructure-activity relationships. New embodiments of1-(4-aminophenyl)pyrazoles, potent and selective for inhibition of sEHare disclosed herein.

[0013] All references cited above and throughout this application areincorporated herein by reference in their entirety.

SUMMARY OF THE INVENTION

[0014] It is therefore an object of the invention to provide a method oftreating a cardiovascular disease; said method comprising administeringto a patient in need thereof a therapeutically effective amount of acompound of Formula I:

[0015] wherein:

[0016] R₁ and R₃ are the same or different and each is CF₃, halogen, CN,C₁₋₈ alkyl or branched alkyl, C₂₋₈ alkenyl or C₃₋₈ branched alkenyl,C₂₋₈ alkynyl or C₃₋₈ branched alkynyl, C₃₋₈ cycloalkyl optionallysubstituted with OH, CN or methoxy, C₁₋₈ alkyloxy, C₁₋₄ alkyloxyC₁₋₄alkyl, C₁₋₈ alkylthio, C₁₋₄ alkylthioC₁₋₄alkyl, C₁₋₈ dialkylamino, C₁₋₄dialkylaminoalkyl, CO₂R₅ where R₅ is C₁₋₄ alkyl or C₂₋₄ alkenyloptionally substituted with carbocyclyl or heterocyclyl, aryl or R₁ andR₃ are heterocyclyl connected to the pyrazole in any position that makesa stable bond optionally substituted with halogen, C₁₋₄ alkyl, C₂₋₄alkenyl, CN, (CH₃)₂N, CO₂CH₃, alkyloxy, aryl, heterocyclyl or R₅;

[0017] R₂ is H, halogen or methyl,

[0018] L is —NHC(O)—, —NHC(O)O—, —NHC(O)C(O)—, —NHC(S)—, —NH—,—NHC(O)NH, NHC(S)NH, NHCH₂, —NHCH(R₆)—, where R₆ is H, CN or C₁₋₃ alkyl,

[0019] R₄ is C₁₋₈ alkyl, C₁₋₈ alkyloxy, C₁₋₈ alkylthio, C₁₋₈ alkylamino,C₁₋₄ alkyloxyalkyl, C₁₋₄ alkylthioalkyl, C₁₋₄alkylaminoalkyl,C₁₋₄dialkylaminoalkyl, carbocyclyl or heterocyclyl each optionallysubstituted with one or more halogen, —CN, —NO₂, SO₂NH₂ alkylthio,alkylsulfinyl, alkylsulfonyl or R₇ where R₇ is phenyl, heterocyclyl,C₃₋₆ cycloalkyl, C₁₋₆ alkyl, C₂₋₆ alkenyl, C₁₋₆ alkyloxyalkyl, C₁₋₄alkyloxy, C₁₋₅ alkylamino, C₁₋₆ alkylthioalkyl, C₁₋₆ alkylsulfinylalkylor C₁₋₆ alkylsulfonylalkyl, each R₇ in turn is optionally substitutedwith halogen, OH, alkyloxy, CN, COO-lower alkyl, —CONH-lower alkyl,—CON(lower alkyl)₂, dialkylamino, phenyl or heterocylcyl;

[0020] R₈ is H or NH₂;

[0021] or the pharmaceutically acceptable derivatives thereof;

[0022] with the proviso that when R₃ is alkyl or CF₃ and R₄ is pyridyl,then the pyridyl is substituted except that the substituents on thepyridyl cannot be halogen; and with the proviso that the followingcompounds are excluded:N-[4-(5-ethyl-3-pyridin-3-yl-pyrazol-1-yl)-phenyl]-nicotinamide;N-[4-(5-Ethyl-3-pyridin-3-yl-pyrazol-1-yl)phenyl]-1-methylindole-2-carboxamide;4-(3-Cyanopropoxy)-N-[4-(5-cyano-3-pyridin-3-yl-pyrazol-1-yl)phenyl]benzamide;andN-[4-(5-cyano-3-pyridin-3-yl-pyrazol-1-yl)phenyl]-4-(3-[1,3]dioxolan-2-yl-propoxy)benzamide.

DETAILED DESCRIPTION OF THE INVENTION

[0023] Preferred embodiments of the invention include:

[0024] The method as described in the broadest embodiment above andwherein:

[0025] in formula (I):

[0026] R₁ is C₁₋₈ alkyl or branched alkyl, C₃₋₈ alkenyl or branchedalkenyl, C₃₋₈ alkynyl or branched alkynyl, C₃₋₈ cycloalkyl, C₁₋₃alkyloxyC₁₋₃ alkyl, C₁₋₅ alkyloxy, C₁₋₃ alkylthioC₁₋₃ alkyl, C₁₋₅alkylthio, CF₃, heterocyclyl selected from tetrahydrofuranyl, pyridyl,furanyl or thiazolyl or aryl optionally substituted with halogen, C₁₋₄alkyl, CN, alkyloxy or (CH₃)₂N;

[0027] R₂ is H;

[0028] R₃ is halogen, methyl, ethyl, CF₃, CN, cyclopropyl, vinyl, SCH₃,methoxy, heterocyclyl selected from tetrahydrofuranyl, pyridyl, furanylor thiazolyl or aryl optionally substituted with halogen, C₁₋₄ alkyl,CN, methoxy or (CH₃)₂N;

[0029] L is —NHC(O)—, —NH—, —NHCH₂—, —NHC(O)NH, and

[0030] R₄ is C₁₋₆ alkyl, carbocyclyl or heterocyclyl selected frompyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, morpholinyl,thiomorpholinyl, pyrrolyl, imidazolyl, pyrazolyl, thienyl, furyl,isoxazolyl, isothiazolyl, oxazolyl, thiazolyl, oxadiazolyl,thiadiazolyl, quinolinyl, isoquinolinyl, indolyl, benzimidazolyl,benzofuranyl, benzoxazolyl, benzisoxazolyl, benzpyrazolyl,benzothiofuranyl, benzothiazolyl, quinazolinyl and indazolyl, eachoptionally substituted with one or more halogen, —CN, alkylthio,alkylsulfinyl, alkylsulfonyl, —NO₂, SO₂NH₂ or R₇ where R₇ is C₁₋₆ alkyl,C₂₋₆ alkenyl, C₁₋₆ alkyloxyalkyl, C₁₋₄ alkyloxy, C₁₋₅ alkylamino, orC₁₋₆ alkylthioalkyl each optionally substituted with OH, CN, —COO-loweralkyl, —CONH-lower alkyl, —CON(lower alkyl)₂, dialkylamino, phenyl orheterocyclyl as hereinabove described in this paragraph; and

[0031] R₈ is H or NH₂.

[0032] In another embodiment, there is provided the method as describedin the embodiment immediately above and wherein:

[0033] in the formula (I)

[0034] R₁ is ethyl, isopropyl, n-propyl, t-butyl, cyclopentyl, CF₃,ethoxy, CH₃OCH₂—, 2- or 3-tetrahydrofuranyl, 2-, 3-, or 4-pyridyl,2-furanyl, or 2-thiazolyl;

[0035] R₃ is CN, CF₃, Cl, methyl, ethyl, SCH₃, cyclopropyl, vinyl or2-furanyl;

[0036] L is —NHC(O)—,

[0037] and

[0038] R₄ is a phenyl or pyridyl each optionally substituted with one tothree halogen, —CN, alkylthio, alkylsulfinyl, alkylsulfonyl or R₇ whereR₇ is C₁₋₆ alkyl, C₂₋₆ alkenyl, C₁₋₆ alkyloxyC₁₋₆ alkyl, C₁₋₄ alkyloxy,C₁₋₅ alkylamino each optionally substituted with halogen, OH, CN,COO-lower alkyl, —CONH-lower alkyl, —CON(lower alkyl)₂, dialkylamino,phenyl, morpholinyl or pyridyl.

[0039] In yet another embodiment, there is provided the method asdescribed in the embodiment immediately above and wherein:

[0040] in the formula (I)

[0041] R₁ is isopropyl, CF₃, 3-pyridyl or 4-pyridyl;

[0042] R₂ is H;

[0043] R₃ is CN, CF₃, Cl, methyl, SCH₃ or ethyl;

[0044] and

[0045] R₄ is a phenyl or pyridyl each optionally substituted with one tothree groups selected from halogen, —CN, alkylthio, alkylsulfinyl,alkylsulfonyl or R₇ where R₇ is C₁₋₆ alkyl, C₁₋₄ alkyloxy, C₁₋₅alkylamino each optionally substituted with OH, CN, COO-lower alkyl,—CONH-lower alkyl, —CON(lower alkyl)₂, dialkylamino, phenyl, morpholinylor pyridyl.

[0046] In yet still another embodiment, there is provided a method oftreating cardiovascular disease said method comprising administering toa patient in need thereof a therapeutically effective amount of acompound chosen from:

[0047] or the pharmaceutically acceptable derivatives thereof.

[0048] In yet still another embodiment, there is provided a method oftreating cardiovascular disease said method comprising administering toa patient in need thereof a therapeutically effective amount of acompound chosen from:

[0049] or the pharmaceutically acceptable derivatives thereof.

[0050] In yet another embodiment of the invention there are providednovel compounds of the formula (Ia)

[0051] wherein:

[0052] R₁ and R₃ are the same or different and each is CF₃, halogen, CN,C₁₋₈ alkyl or branched alkyl, C₂₋₈ alkenyl or C₃₋₈ branched alkenyl,C₂₋₈ alkynyl or C₃₋₈ branched alkynyl, C₃₋₈ cycloalkyl optionallysubstituted with OH, CN or methoxy, C₁₋₈ alkyloxy, C₁₋₄ alkyloxyC₁₋₄alkyl, C₁₋₈ alkylthio, C₁₋₄ alkylthioC₁₋₄alkyl, C₁₋₈ dialkylamino, C₁₋₄dialkylaminoalkyl, CO₂R₅ where R₅ is C₁₋₄ alkyl or C2-4 alkenyloptionally substituted with carbocyclyl or heterocyclyl, aryl or R₁ andR₃ are heterocyclyl connected to the pyrazole in any position that makesa stable bond optionally substituted with halogen, C₁₋₄ alkyl, C₂₋₄alkenyl, CN, (CH₃)₂N, CO₂CH₃, alkyloxy, aryl, heterocyclyl or R₅;

[0053] R₂ is H, halogen or methyl;

[0054] L is —NHC(O)—, —NHC(O)O—, —NHC(O)C(O)—, —NHC(S)—, —NH—,—NHC(O)NH, NHC(S)NH, NHCH₂, —NHCH(R₆)—, where R₆ is H, CN or C₁₋₃ alkyl,

[0055] R₄ is C₁₋₈ alkyl, C₁₋₈ alkyloxy, C₁₋₈ alkylthio, C₁₋₈ alkylamino,C₁₋₄ alkyloxyalkyl, C₁₋₄ alkylthioalkyl, C₁₋₄alkylaminoalkyl,C₁₋₄dialkylaminoalkyl, carbocyclyl or heterocyclyl each optionallysubstituted with one or more halogen, —CN, —NO₂, SO₂NH₂ alkylthio,alkylsulfinyl, alkylsulfonyl or R₇ where R₇ is phenyl, heterocyclyl,C₃₋₆ cycloalkyl, C₁₋₆ alkyl, C₂₋₆ alkenyl, C₁₋₆ alkyloxyalkyl, C₁₋₄alkyloxy, C₁₋₅ alkylamino, C₁₋₆ alkylthioalkyl, C₁₋₆ alkylsulfinylalkylor C₁₋₆ alkylsulfonylalkyl, each R₇ in turn is optionally substitutedwith halogen, OH, alkyloxy, CN, COO-lower alkyl, —CONH-lower alkyl,—CON(lower alkyl)₂, dialkylamino, phenyl or heterocylcyl;

[0056] R₈ is NH₂ or mono-or-diC1-5alkylamino;

[0057] or the pharmaceutically acceptable derivatives thereof.

[0058] Preferred embodiments of the formula(Ia) include:

[0059] The compound of the formula(Ia) as described in the broadestembodiment above and wherein:

[0060] R₁ is C₁₋₈ alkyl or branched alkyl, C₃₋₈ alkenyl or branchedalkenyl, C₃₋₈ alkynyl or branched alkynyl, C₃₋₈ cycloalkyl, C₁₋₃alkyloxyC₁₋₃ alkyl, C₁₋₅ alkyloxy, C₁₋₃ alkylthioC₁₋₃ alkyl, C₁₋₅alkylthio, CF₃, heterocyclyl selected from tetrahydrofuranyl, pyridyl,furanyl or thiazolyl or aryl optionally substituted with halogen, C₁₋₄alkyl, CN, alkyloxy or (CH₃)₂N;

[0061] R₂ is H;

[0062] R₃ is halogen, methyl, ethyl, CF₃, CN, cyclopropyl, vinyl, SCH₃,methoxy, heterocyclyl selected from tetrahydrofuranyl, pyridyl, furanylor thiazolyl or aryl optionally substituted with halogen, C₁₋₄ alkyl,CN, methoxy or (CH₃)₂N;

[0063] L is —NHC(O)—, —NH—, —NHCH₂—, —NHC(O)NH, and

[0064] R₄ is C₁₋₆ alkyl, carbocyclyl or heterocyclyl selected frompyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, morpholinyl,thiomorpholinyl, pyrrolyl, imidazolyl, pyrazolyl, thienyl, furyl,isoxazolyl, isothiazolyl, oxazolyl, thiazolyl, oxadiazolyl,thiadiazolyl, quinolinyl, isoquinolinyl, indolyl, benzimidazolyl,benzofuranyl, benzoxazolyl, benzisoxazolyl, benzpyrazolyl,benzothiofuranyl, benzothiazolyl, quinazolinyl and indazolyl, eachoptionally substituted with one or more halogen, —CN, alkylthio,alkylsulfinyl, alkylsulfonyl , —NO₂, SO₂NH₂ or R₇ where R₇ is C₁₋₆alkyl, C₂₋₆ alkenyl, C₁₋₆ alkyloxyalkyl, C₁₋₄ alkyloxy, C₁₋₅ alkylamino,or C₁₋₆ alkylthioalkyl each optionally substituted with OH, CN,—COO-lower alkyl, —CONH-lower alkyl, —CON(lower alkyl)₂, dialkylamino,phenyl or heterocyclyl as hereinabove described in this paragraph; and

[0065] R₈ is NH₂.

[0066] In another embodiment, there is provided compounds of theformula(Ia) as described in the embodiment immediately above andwherein:

[0067] R₁ is ethyl, isopropyl, n-propyl, t-butyl, cyclopentyl, CF₃,ethoxy, CH₃OCH₂—, 2- or 3-tetrahydrofuranyl, 2-, 3-, or 4-pyridyl,2-furanyl, or 2-thiazolyl;

[0068] R₃ is CN, CF₃, Cl, methyl, ethyl, SCH₃, cyclopropyl, vinyl or2-furanyl;

[0069] L is —NHC(O)—,

[0070] and

[0071] R₄ is a phenyl or pyridyl each optionally substituted with one tothree halogen, —CN, alkylthio, alkylsulfinyl, alkylsulfonyl or R₇ whereR₇ is C₁₋₆ alkyl, C₂₋₆ alkenyl, C₁₋₆ alkyloxyC₁₋₆ alkyl, C₁₋₄ alkyloxy,C₁₋₅ alkylamino each optionally substituted with halogen, OH, CN,COO-lower alkyl, —CONH-lower alkyl, —CON(lower alkyl)₂, dialkylamino,phenyl, morpholinyl or pyridyl.

[0072] In yet another embodiment, there is provided compounds of theformula(Ia) as described in the embodiment immediately above andwherein:

[0073] R₁ is isopropyl, CF₃, 3-pyridyl or 4-pyridyl;

[0074] R₂ is H;

[0075] R₃ is CN, CF₃, Cl, methyl, SCH₃ or ethyl;

[0076] and

[0077] R₄ is a phenyl or pyridyl each optionally substituted with one tothree groups selected from halogen, —CN, alkylthio, alkylsulfinyl,alkylsulfonyl or R₇ where R₇ is C₁₋₆ alkyl, C₁₋₄ alkyloxy, C₁₋₅alkylamino each optionally substituted with OH, CN, COO-lower alkyl,—CONH-lower alkyl, —CON(lower alkyl)₂, dialkylamino, phenyl, morpholinylor pyridyl.

[0078] A particularly preferred embodiment of formula Ia is

[0079] Any of the of compounds of formulas I or Ia containing one ormore asymmetric carbon atoms may occur as racemates and racemicmixtures, single enantiomers, diastereomeric mixtures and individualdiastereomers. All such isomeric forms of these compounds are expresslyincluded in the present invention. Each stereogenic carbon may be in theR or S configuration, or a combination of configurations.

[0080] Some of the compounds of formulas I or Ia can exist in more thanone tautomeric form. The invention includes use of all such tautomers.

[0081] The compounds of formulas I or Ia are only those which arecontemplated to be ‘chemically stable’ as will be appreciated by thoseskilled in the art. For example, compounds which would have a ‘danglingvalency’, or a ‘carbanion’ are not compounds contemplated to be used inthe methods of the invention.

[0082] All terms as used herein in this specification, unless otherwisestated, shall be understood in their ordinary meaning as known in theart. All alkyl, alkylene, alkenyl, alkenylene, alkynyl and alkynylenegroups shall be understood as being C₁₋₁₀, branched or unbranched unlessotherwise specified. Other more specific definitions are as follows:

[0083] A “pharmaceutically acceptable derivative” refers to anypharmaceutically acceptable salt or ester of a compound of thisinvention, or any other compound which, upon administration to apatient, is capable of providing (directly or indirectly) a compoundused in this invention, a pharmacologically active metabolite orpharmacologically active residue thereof.

[0084] The term “metabolite” shall be understood to mean any of thecompounds of the formula I or Ia which are capable of being hydroxylatedor oxidized, enzymatically or chemically, as will be appreciated bythose skilled in the art.

[0085] The term “acyl”, when used alone or in combination with anothergroup, shall be understood to mean an R—(C═O)-moiety wherein R is analkyl group. Examples of R can be a C₁₋₁₀alkyl, saturated orunsaturated, branched or unbranched. The term “acyloxy” shall beunderstood to mean an R—CO₂-group wherein R is as defined in thisparagraph. Likewise, “acylthio” shall be understood to mean anR—C(O)—S-group wherein R is as defined in this paragraph. “Alkyloxy”shall be understood to mean an R—O— group wherein R is as defined inthis paragraph

[0086] The term “alkylene” shall be understood to mean a saturated,divalent C₁₋₁₀ hydrocarbon chain, i.e., generally present as a bridginggroup between two other groups. Examples of alkylene groups include—CH₂-(methylene); —CH₂CH₂-(ethylene); —CH₂CH₂CH₂-(propylene), etc.

[0087] The term “alkenylene” shall be understood to mean a divalentC₁₋₁₀ hydrocarbon chain having one or more double bonds within thechain, i.e., generally present as a bridging group between two othergroups. Examples of alkenylene groups include —CH═CH— (ethenylene);—CH═CHCH₂-(1-propenylene), —CH═CHCH₂CH₂-(1-butenylene),—CH₂CH═CHCH₂-(2-butenylene), etc.

[0088] The term “alkynylene” shall be understood to mean a divalentC₁₋₁₀ hydrocarbon chain having one or more triple bonds within thechain, i.e., generally present as a bridging group between two othergroups. Examples of alkenylene groups include 1'C≡C—; —C≡CCH₂—;—C≡CCH₂CH₂—; —CH₂C≡CCH₂—, etc.

[0089] The term “aryl” shall be understood to mean a 6-10 memberedaromatic carbocycle; “aryl” includes, for example, phenyl and naphthyl;other terms comprising “aryl” will have the same definition for the arylcomponent, examples of these moieties include: arylalkyl, aryloxy orarylthio.

[0090] The term “cycloalkenyl” shall be understood to mean aC₃₋₁₀cycloalkyl group wherein one or more of the single bonds in thecycloalkyl ring are replaced by double bonds.

[0091] The terms “cycloalkylene” and “cycloalkenylene” shall beunderstood to mean divalent C₄₋₁₀cycloalkyl and C₄₋₁₀cycloalkenylgroups, respectively, i.e., generally present as bridging groups betweentwo other groups.

[0092] The term “halogen” as used in the present specification shall beunderstood to mean bromine, chlorine, fluorine or iodine.

[0093] The term “heteroaryl” refers to a stable 5-8 membered (butpreferably, 5 or 6 membered) monocyclic or 8-11 membered bicyclicaromatic heterocycle radical. Each heterocycle consists of carbon atomsand from 1 to 4 heteroatoms chosen from nitrogen, oxygen and sulfur. Theheterocycle may be attached by any atom of the cycle, which results inthe creation of a stable structure. Example “heteroaryl” radicalsinclude, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, pyrrolyl,imidazolyl, pyrazolyl, thienyl, furyl, isoxazolyl, isothiazolyl,oxazolyl, thiazolyl, oxadiazolyl, thiadiazolyl, quinolinyl,isoquinolinyl, indolyl, benzimidazolyl, benzofuranyl, benzoxazolyl,benzisoxazolyl, benzpyrazolyl, benzothiofuranyl, benzothiazolyl,quinazolinyl and indazolyl,or a fused heteroaryl such ascyclopentenopyridine, cyclohexanopyridine, cyclopentanopyrimidine,cyclohexanopyrimidine, cyclopentanopyrazine, cyclohexanopyrazine,cyclopentanopyridazine, cyclohexanopyridazine, cyclopentanoquinoline,cyclohexanoquinoline, cyclopentanoisoquinoline, cyclohexanoisoquinoline,cyclopentanoindole, cyclohexanoindole, cyclopentanobenzimidazole,cyclohexanobenzimidazole, cyclopentanobenzoxazole,cyclohexanobenzoxazole, cyclopentanoimidazole, cyclohexanoimidazole,cyclopentanothiophene and cyclohexanothiophene;

[0094] The term “heterocycle” refers to a stable 5-8 membered (butpreferably, 5 or 6 membered) monocyclic or 8-11 membered bicyclicheterocycle radical which may be either saturated or unsaturated, and isnon-aromatic. Each heterocycle consists of carbon atoms and from 1 to 4heteroatoms chosen from nitrogen, oxygen and sulfur. The heterocycle maybe attached to the main structure by any atom of the cycle, whichresults in the creation of a stable structure. Example “heterocycle”radicals include pyrrolinyl, pyrrolidinyl, pyrazolinyl, pyrazolidinyl,1,2,5,6-tetrahydropyridinyl, piperidinyl, morpholinyl, thiomorpholinyl,pyranyl, thiopyranyl, piperazinyl, indolinyl, and1,2,3,3a,4,6a-hexahydro-cyclopenta[c]pyrrolyl.

[0095] As used herein and throughout this specification, the terms“nitrogen” and “sulfur” and their respective elements symbols includeany oxidized form of nitrogen and sulfur and the quaternized form of anybasic nitrogen.

[0096] The “C₆₋₁₂ bridged carbocyclic ring system, optionally having oneto three double bonds in the ring system” shall be understood to meanany carbocyclic ring system containing 6 to 12 carbon atoms and havingat least one bridged-type fusion within the ring system. An example is aC₆₋₁₀carbocyclic ring system, optionally having one or two double bondsin the system. Examples of such a ring system are bicyclo[2.2.1]heptaneand adamantane.

[0097] Methods of making all compounds described herein are thosemethods well known in the art and in particular those described in WO99/62885, and the cited methods therein, are incorporated herein byreference in their entirety.

[0098] In order that this invention be more fully understood, thefollowing examples are set forth. These examples are for the purpose ofillustrating preferred embodiments of this invention, and are not to beconstrued as limiting the scope of the invention in any way.

[0099] The examples which follow are illustrative and, as recognized byone skilled in the art, particular reagents or conditions could bemodified as needed for individual compounds without undueexperimentation. Starting materials used in the scheme below are eithercommercially available or easily prepared from commercially availablematerials by those skilled in the art.

METHODS OF USE

[0100] In accordance with the invention, there are provided methods ofusing the compounds of the formulas I or Ia. The compounds used in theinvention prevent the degradation of sEH substrates that have beneficialeffects or prevent the formation of metabolites that have adverseeffects. The inhibition of sEH is an attractive means for preventing andtreating a variety of cardiovascular diseases or conditions e.g.,endothelial dysfunction. Thus, the methods of the invention are usefulfor the treatment of such conditions. These encompass diseasesincluding, but not limited to, type 1 and type 2 diabetes, insulinresistance syndrome, hypertension, atherosclerosis, coronary arterydisease, angina, ischemia, ischemic stroke, Raynaud's disease and renaldisease.

[0101] For therapeutic use, the compounds may be administered in anyconventional dosage form in any conventional manner. Routes ofadministration include, but are not limited to, intravenously,intramuscularly, subcutaneously, intrasynovially, by infusion,sublingually, transdermally, orally, topically or by inhalation. Thepreferred modes of administration are oral and intravenous.

[0102] The compounds described herein may be administered alone or incombination with adjuvants that enhance stability of the inhibitors,facilitate administration of pharmaceutic compositions containing themin certain embodiments, provide increased dissolution or dispersion,increase inhibitory activity, provide adjunct therapy, and the like,including other active ingredients. Advantageously, such combinationtherapies utilize lower dosages of the conventional therapeutics, thusavoiding possible toxicity and adverse side effects incurred when thoseagents are used as monotherapies. Compounds of the invention may bephysically combined with the conventional therapeutics or otheradjuvants into a single pharmaceutical composition. Advantageously, thecompounds may then be administered together in a single dosage form. Insome embodiments, the pharmaceutical compositions comprising suchcombinations of compounds contain at least about 5%, but more preferablyat least about 20%, of a compound of formula (I) (w/w) or a combinationthereof. The optimum percentage (w/w) of a compound of the invention mayvary and is within the purview of those skilled in the art.Alternatively, the compounds may be administered separately (eitherserially or in parallel). Separate dosing allows for greater flexibilityin the dosing regime.

[0103] As mentioned above, dosage forms of the above-described compoundsinclude pharmaceutically acceptable carriers and adjuvants known tothose of ordinary skill in the art. These carriers and adjuvantsinclude, for example, ion exchangers, alumina, aluminum stearate,lecithin, serum proteins, buffer substances, water, salts orelectrolytes and cellulose-based substances. Preferred dosage formsinclude, tablet, capsule, caplet, liquid, solution, suspension,emulsion, lozenges, syrup, reconstitutable powder, granule, suppositoryand transdermal patch. Methods for preparing such dosage forms are known(see, for example, H. C. Ansel and N. G. Popovish, Pharmaceutical DosageForms and Drug Delivery Systems, 5th ed., Lea and Febiger (1990)).Dosage levels and requirements are well-recognized in the art and may beselected by those of ordinary skill in the art from available methodsand techniques suitable for a particular patient. In some embodiments,dosage levels range from about 1-1000 mg/dose for a 70 kg patient.Although one dose per day may be sufficient, up to 5 doses per day maybe given. For oral doses, up to 2000 mg/day may be required. As theskilled artisan will appreciate, lower or higher doses may be requireddepending on particular factors. For instance, specific dosage andtreatment regimens will depend on factors such as the patient's generalhealth profile, the severity and course of the patient's disorder ordisposition thereto, and the judgment of the treating physician.

EXAMPLES Example I

[0104]

[0105] To a solution of 2-chloroniotinic acid (0.78 g) in acetonitrile(25 mL) cooled on ice was added EDC (1.1 g). After 10 minutes, theaniline I (1.0 g) was added. The mixture was stirred on ice for 1 hourand then allowed to warm to room temperature. The solid product II wascollected by filtration (1.2 g).

[0106] A mixture of II (0.1 g) and 5-amino pentanol (0.047 g) in dioxane(2 mL) was heated at 120° C. in a sealed tube, overnight. The mixturewas cooled, diluted with ethyl acetate, washed with water, dried,filtered and evaporated. Purification by preparative layerchromatography gave III as a solid (0.05 g), mp 95-96° C.

[0107] A mixture of II (0.1 g) and benzylamine (1.5 mL) was heated in asealed tube at 120° C. overnight. The mixture was cooled, diluted withmethylene chloride, washed with water, dried filtered and evaporated.Chromatography of the residue over silica gel gave IV (0.06 g), mp183-184° C.

[0108] A mixture of II (0.1 g) and aminoethylmorpholine (0.059 g) indioxane (2 mL) was heated at 120° C. in a sealed tube, overnight. Themixture was diluted with ethyl acetate, washed with water, dried,filtered and evaporated. Chromatography of the residue over silica gelgave V (0.045 g) mp 85-87° C.

[0109] To a solution of VI (0.3 g) in methylene chloride (5 mL) wasadded diisopropylethylamine (0.18 mL) followed by 4-morpholine carbonylchloride (0.11 mL). The mixture was stirred at room temperature for 3days. The mixture was diluted with ethyl acetate, washed with water,dried, filtered and evaporated. Chromatography of the residue oversilica gel gave VII (0.8 g) mp 185-186° C.

[0110] To a solution of 2-thiomethyl nicotinic acid (0.217 g) inacetonitrile cooled on ice, was added EDC (0.27 g). After 10 minutes, I(0.30 g) was added along with DMAP (catalytic amount). The mixture wasstirred on ice for 1 hour and then allowed to stir at room temperatureovernight. The mixture was diluted with ethyl acetate, washed withwater, dried filtered and evaporated to give (VIII).

[0111] To a solution of VIII (0.09 g) in methylene chloride (7 mL)cooled on ice, was added m-chloroperbenzoic acid (0.043 g). After 10minutes, the reaction mixture was washed with aqueous sodiumbicarbonate, dried, filtered and evaporated. Purification by preparativelayer chromatography gave (IX) (0.017 g), mp 243-244° C.

[0112] Compounds with R₈═NH₂ or mono-or-dialkylamino, may be prepared bythe methods described below.

[0113] A 3,5-disubstituted pyrazole may be reacted with dinitrobenzenesubstituted in the 4-position with a leaving group such as a halogen inthe presence of a base. The nitrophenylpyrazoles produced by eithermethod could then be reduced to diaminophenyl pyrazoles by using areducing agent such as SnCl₂ or hydrogen or a hydrogen source such asammonium formate in the presence of a catalyst such as palladium. Thediamino compound could then be converted to compounds of Formula 1(R₈═NH₂, or mono-or-dialkylamino) by methods previously described.

[0114] Alternatively, a compound of Formula I (where R₈═H) can bereacted with a nitrating reagent such as nitronium tetrafluoroborate toprovide the nitrated intermediate shown above. Reduction of the nitrogroup by using a reducing agent such as SnCl₂ or hydrogen or a hydrogensource such as ammonium formate in the presence of a catalyst such aspalladium then provides the compounds of formula I/Ia where R₈═NH₂.Alkylation of the NH₂ by methods known in the art provides compoundswith R₈=mono, dialkylamine. For example treatment with formaldehyde andformic acid results in R₈═NMe₂.

Example II

[0115]

[0116] To a solution of X (0.65 g) in acetonitrile (10 mL) and methylenechloride (3 mL) cooled on ice was added nitronium tetrafluoroborate(0.575 g) in two portions over 10 minutes. After 10 minutes, thereaction was quenched by addition of saturated aqueous sodiumbicarbonate. Ethyl acetate was added and the organic phase was washedwith water, dried, filtered and evaporated. Crystallization of theproduct from ethanol/methylene chloride gave XI (0.597 g).

[0117] To a solution of XI (0.22 g) in acetic acid (10 mL) was added asolution of stannous chloride (0.73 g) in cone. HCl (5 mL). Additionalacetic acid (5 mL) was added and the mixture was stirred at roomtemperature for 6 hours. The mixture was neutralized with aqueous KOHand extracted with methylene chloride. The organic phase was dried,filtered and evaporated.to give XII (0.18 g) , mp 201-203° C.

[0118] Preferred embodiments of the invention include methods of usingparticular inhibitors which have been found to be surprisingly effectiveat inhibiting the sEH enzyme. Methods employed for selecting suchinhibitors include the fluorescence polarization assay summarized belowand are described in U.S. provisional application serial No. 60/282,575,incorporated herein by reference in it's entirety.

[0119] Fluorescence Polarization Assay to Determine Inhibition of sEH:

[0120] Step one: Characterization of the Fluorescent Probe

[0121] The wavelengths for maximum excitation and emission of thefluorescent probe should first be measured. An example of such a probeis compound (4) as shown in U.S. Ser. No. 60/282,575, where these valuesare 529 nm and 565 nm, respectively. These fluorescence wavelengthvalues were measured on an SLM-8100 fluorimeter with the probe dissolvedin an assay buffer (20 mM TES, pH 7.0, 200 mM NaCl, 0.05% (w/v) CHAPS, 2mM DTT).

[0122] The affinity of the probe for sEH was then determined in atitration experiment. The fluorescence polarization value of compound 4in assay buffer was measured on an SLM-8100 fluorimeter using theexcitation and emission maximum values described above. Aliquots of sEHwere added and fluorescence polarization was measured after eachaddition until no further change in polarization value was observed.Non-linear least squares regression analysis was used to calculate thedissociation constant of compound 4 from the polarization valuesobtained for sEH binding to compound 4. FIG. 1 shows the results fromthis titration experiment

[0123] Step two: Screening for Inhibitors of Probe Binding

[0124] In order to screen a large number of compounds the assay wasperformed using a 96-well plate format. An example of such a plate isthe Dynex Microfluor 1, low protein binding U-bottom black 96 wellplates (# 7005). The plate is set up by first creating a complex betweenrecombinant human sEH and a fluorescent probe that binds to the activesite of sEH. In this example, the complex between compound 4 and sEH,was pre-formed in assay buffer (20 mM TES, pH 7.0, 200 mM NaCl, 0.05%(w/v) CHAPS, 1 mM TCEP). The concentrations of sEH and compound 4 inthis solution were made up such that the final concentration in theassay was 10 nM sEH and 2.5 nM compound 4. Test compounds were thenserially diluted into assay buffer, across a 96 well plate .Thepre-formed sEH-probe complex was then added to all the wells andincubated for 15 minutes at room temperature. The fluorescencepolarization was then measured using a fluorescence polarization platereader set at the wavelengths appropriate for the fluorescent label onthe fluorescent probe (4). In this example, an LJL Analyst was set toread rhodamine fluorescence polarization (Ex 530 nM, Em 580 nM) .Non-linear least squares regression analysis was then used to calculatedissociation constants for the test compounds binding to sEH from thepolarization values for the probe binding to sEH in the presence of thetest compounds.

[0125] Results which show a decrease in fluorescence polarization of theprobe-sEH complex in the presence of the test compound is evidence thatthis test compound is a competitive inhibitor of soluble epoxidehydrolase that competes with the fluorescent probe for sEH active sitebinding.

What is claimed is:
 1. A method of treating a cardiovascular disease,said method comprising administering to a patient in need thereof atherapeutically effective amount of a compound of Formula I:

wherein: R₁ and R₃ are the same or different and each is CF₃, halogen,CN, C₁₋₈ alkyl or branched alkyl, C₂₋₈ alkenyl or C₃₋₈ branched alkenyl,C₂₋₈ alkynyl or C₃₋₈ branched alkynyl, C₃₋₈ cycloalkyl optionallysubstituted with OH, CN or methoxy, C₁₋₈ alkyloxy, C₁₋₄alkyloxyC₁₋₄alkyl, C₁₋₈ alkylthio, C₁₋₄ alkylthioC₁₋₄alkyl, C₁₋₈dialkylamino, C₁₋₄ dialkylaminoalkyl, CO₂R₅ where R₅ is C₁₋₄ alkyl orC₂₋₄ alkenyl optionally substituted with carbocyclyl or heterocyclyl,aryl or R₁ and R₃ are heterocyclyl connected to the pyrazole in anyposition that makes a stable bond optionally substituted with halogen,C₁₋₄ alkyl, C₂₋₄ alkenyl, CN, (CH₃)₂N, CO₂CH₃, alkyloxy, aryl,heterocyclyl or R₅; R₂ is H, halogen or methyl; L is —NHC(O)—,—NHC(O)O—, —NHC(O)C(O)—, —NHC(S)—, —NH—, —NHC(O)NH, NHC(S)NH, NHCH₂,—NHCH(R₆)—, where R₆ is H, CN or C₁₋₃ alkyl, R₄ is C₁₋₈ alkyl, C₁₋₈alkyloxy, C₁₋₈ alkylthio, C₁₋₈ alkylamino, C₁₋₄ alkyloxyalkyl, C₁₋₄alkylthioalkyl, C₁₋₄alkylaminoalkyl, C₁₋₄dialkylaminoalkyl, carbocyclylor heterocyclyl each optionally substituted with one or more halogen,—CN, —NO₂, SO₂NH₂ alkylthio, alkylsulfinyl, alkylsulfonyl or R₇ where R₇is phenyl, heterocyclyl, C₃₋₆ cycloalkyl, C₁₋₆ alkyl, C₂₋₆ alkenyl, C₁₋₆alkyloxyalkyl, C₁₋₄ alkyloxy, C₁₋₅ alkylamino, C₁₋₆ alkylthioalkyl, C₁₋₆alkylsulfinylalkyl or C₁₋₆ alkylsulfonylalkyl, each R₇ in turn isoptionally substituted with halogen, OH, alkyloxy, CN, COO-lower alkyl,—CONH-lower alkyl, —CON(lower alkyl)₂, dialkylamino, phenyl orheterocylcyl; R₈ is H or NH₂; or the pharmaceutically acceptablederivatives thereof; with the proviso that when R₃ is alkyl or CF₃ andR₄ is pyridyl, then the pyridyl is substituted except that thesubstituents on the pyridyl cannot be halogen; and with the proviso thatthe following compounds are excluded:N-[4-(5-ethyl-3-pyridin-3-yl-pyrazol-1-yl)-phenyl]-nicotinamide;N-[4-(5-Ethyl-3-pyridin-3-yl-pyrazol-1-yl)phenyl]-1-methylindole-2-carboxamide;4-(3-Cyanopropoxy)-N-[4-(5-cyano-3-pyridin-3-yl-pyrazol-1-yl)phenyl]benzamide;andN-[4-(5-cyano-3-pyridin-3-yl-pyrazol-1-yl)phenyl]-4-(3-[1,3]dioxolan-2-yl-propoxy)benzamide.2. The method according to claim 1 and wherein: in formula (I): R₁ isC₁₋₈ alkyl or branched alkyl, C₃₋₈ alkenyl or branched alkenyl, C₃₋₈alkynyl or branched alkynyl, C₃₋₈ cycloalkyl, C₁₋₃ alkyloxyC₁₋₃ alkyl,C₁₋₅ alkyloxy, C₁₋₃ alkylthioC₁₋₃ alkyl, C₁₋₅ alkylthio, CF₃,heterocyclyl selected from tetrahydrofuranyl, pyridyl, furanyl orthiazolyl or aryl optionally substituted with halogen, C₁₋₄ alkyl, CN,alkyloxy or (CH₃)₂N; R₂ is H; R₃ is halogen, methyl, ethyl, CF₃, CN,cyclopropyl, vinyl, SCH₃, methoxy, heterocyclyl selected fromtetrahydrofuranyl, pyridyl, furanyl or thiazolyl or aryl optionallysubstituted with halogen, C₁₋₄ alkyl, CN, methoxy or (CH₃)₂N; L is—NHC(O)—, —NH—, —NHCH₂—, —NHC(O)NH, and R₄ is C₁₋₆ alkyl, carbocyclyl orheterocyclyl selected from pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl,morpholinyl, thiomorpholinyl, pyrrolyl, imidazolyl, pyrazolyl, thienyl,furyl, isoxazolyl, isothiazolyl, oxazolyl, thiazolyl, oxadiazolyl,thiadiazolyl, quinolinyl, isoquinolinyl, indolyl, benzimidazolyl,benzofuranyl, benzoxazolyl, benzisoxazolyl, benzpyrazolyl,benzothiofuranyl, benzothiazolyl, quinazolinyl and indazolyl, eachoptionally substituted with one or more halogen, —CN, alkylthio,alkylsulfinyl, alkylsulfonyl, —NO₂, SO₂NH₂ or R₇ where R₇ is C₁₋₆ alkyl,C₂₋₆ alkenyl, C₁₋₆ alkyloxyalkyl, C₁₋₄ alkyloxy, C₁₋₅ alkylamino, orC₁₋₆ alkylthioalkyl each optionally substituted with OH, CN, —COO-loweralkyl, —CONH-lower alkyl, —CON(lower alkyl)₂, dialkylamino, phenyl orheterocyclyl as hereinabove described in this paragraph; and R₈ is H orNH₂.
 3. The method according to claim 2 and wherein: in the formula (I)R₁ is ethyl, isopropyl, n-propyl, t-butyl, cyclopentyl, CF₃, ethoxy,CH₃OCH₂—, 2- or 3-tetrahydrofuranyl, 2-, 3-, or 4-pyridyl, 2-furanyl, or2-thiazolyl; R₃ is CN, CF₃, Cl, methyl, ethyl, SCH₃, cyclopropyl, vinylor 2-furanyl; L is —NHC(O)—, and R₄ is a phenyl or pyridyl eachoptionally substituted with one to three halogen, —CN, alkylthio,alkylsulfinyl, alkylsulfonyl or R₇ where R₇ is C₁₋₆ alkyl, C₂₋₆ alkenyl,C₁₋₆ alkyloxyC₁₋₆ alkyl, C₁₋₄ alkyloxy, C₁₋₅ alkylamino each optionallysubstituted with halogen, OH, CN, COO-lower alkyl, —CONH-lower alkyl,—CON(lower alkyl)₂, dialkylamino, phenyl, morpholinyl or pyridyl.
 4. Themethod according to claim 3 and wherein: in the formula (I) R₁ isisopropyl, CF₃, 3-pyridyl or 4-pyridyl; R₂ is H; R₃ is CN, CF₃, Cl,methyl, SCH₃ or ethyl; and R₄ is a phenyl or pyridyl each optionallysubstituted with one to three groups selected from halogen, —CN,alkylthio, alkylsulfinyl, alkylsulfonyl or R₇ where R₇ is C₁₋₆ alkyl,C₁₋₄ alkyloxy, C₁₋₅ alkylamino each optionally substituted with OH, CN,COO-lower alkyl, —CONH-lower alkyl, —CON(lower alkyl)₂, dialkylamino,phenyl, morpholinyl or pyridyl.
 5. A method of treating a cardiovasculardisease, said method comprising administering to a patient in needthereof a therapeutically effective amount of a compound chosen from:

or the pharmaceutically acceptable derivatives thereof.
 6. The methodaccording to claim 5 wherein the compound is chosen from:

or the pharmaceutically acceptable derivatives thereof.
 7. The methodaccording to claims 1 or 5 wherein the cardiovascular disease isinvolves endothelial dysfunction.
 8. The method according to claims 1 or5 wherein the cardiovascular disease is chosen from type 1 and type 2diabetes, insulin resistance syndrome, hypertension, atherosclerosis,coronary artery disease, angina, ischemia, ischemic stroke, Raynaud'sdisease and renal disease.
 9. A compound of the formula (Ia)

wherein: R₁ and R₃ are the same or different and each is CF₃, halogen,CN, C₁₋₈ alkyl or branched alkyl, C₂₋₈ alkenyl or C₃₋₈ branched alkenyl,C₂₋₈ alkynyl or C₃₋₈ branched alkynyl, C₃₋₈ cycloalkyl optionallysubstituted with OH, CN or methoxy, C₁₋₈ alkyloxy, C₁₋₄ alkyloxyC₁₋₄alkyl, C₁₋₈ alkylthio, C₁₋₄ alkylthioC₁₋₄alkyl, C₁₋₈ dialkylamino, C₁₋₄dialkylaminoalkyl, CO₂R₅ where R₅ is C₁₋₄ alkyl or C₂₋₄ alkenyloptionally substituted with carbocyclyl or heterocyclyl, aryl or R₁ andR₃ are heterocyclyl connected to the pyrazole in any position that makesa stable bond optionally substituted with halogen, C₁₋₄ alkyl, C₂₋₄alkenyl, CN, (CH₃)₂N, CO₂CH₃, alkyloxy, aryl, heterocyclyl or R₅; R₂ isH, halogen or methyl; L is —NHC(O)—, —NHC(O)O—, —NHC(O)C(O)—, —NHC(S)—,—NH—, —NHC(O)NH, NHC(S)NH, NHCH₂, —NHCH(R₆)—, where R₆ is H, CN or C₁₋₃alkyl, R₄ is C₁₋₈ alkyl, C₁₋₈ alkyloxy, C₁₋₈ alkylthio, C₁₋₈ alkylamino,C₁₋₄ alkyloxyalkyl, C₁₋₄ alkylthioalkyl, C₁₋₄alkylaminoalkylC₁₋₄dialkylaminoalkyl, carbocyclyl or heterocyclyl each optionallysubstituted with one or more halogen, —CN, —NO₂, SO₂NH₂ alkylthio,alkylsulfinyl, alkylsulfonyl or R₇ where R₇ is phenyl, heterocyclyl,C₃₋₆ cycloalkyl, C₁₋₆ alkyl, C₂₋₆ alkenyl, C₁₋₆ alkyloxyalkyl, C₁₋₄alkyloxy, C₁₋₅ alkylamino, C₁₋₆ alkylthioalkyl, C₁₋₆ alkylsulfinylalkylor C₁₋₆ alkylsulfonylalkyl, each R₇ in turn is optionally substitutedwith halogen, OH, alkyloxy, CN, COO-lower alkyl, —CONH-lower alkyl,—CON(lower alkyl)₂, dialkylamino, phenyl or heterocylcyl; R₈ is NH₂ ormono-or-diC1-5alkylamino; or the pharmaceutically acceptable derivativesthereof.
 10. The compound according to claim 9 wherein: R₁ is C₁₋₈ alkylor branched alkyl, C₃₋₈ alkenyl or branched alkenyl, C₃₋₈ alkynyl orbranched alkynyl, C₃₋₈ cycloalkyl, C₁₋₃ alkyloxyC₁₋₃ alkyl, C₁₋₅alkyloxy, C₁₋₃ alkylthioC₁₋₃ alkyl, C₁₋₅ alkylthio, CF₃, heterocyclylselected from tetrahydrofuranyl, pyridyl, furanyl or thiazolyl or aryloptionally substituted with halogen, C₁₋₄ alkyl, CN, alkyloxy or(CH₃)₂N; R₂ is H; R₃ is halogen, methyl, ethyl, CF₃, CN, cyclopropyl,vinyl, SCH₃, methoxy, heterocyclyl selected from tetrahydrofuranyl,pyridyl, furanyl or thiazolyl or aryl optionally substituted withhalogen, C₁₋₄ alkyl, CN, methoxy or (CH₃)₂N; L is —NHC(O)—, —NH—,—NHCH₂—, —NHC(O)NH, and R₄ is C₁₋₆ alkyl, carbocyclyl or heterocyclylselected from pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, morpholinyl,thiomorpholinyl, pyrrolyl, imidazolyl, pyrazolyl, thienyl, furyl,isoxazolyl, isothiazolyl, oxazolyl, thiazolyl, oxadiazolyl,thiadiazolyl, quinolinyl, isoquinolinyl, indolyl, benzimidazolyl,benzofuranyl, benzoxazolyl, benzisoxazolyl, benzpyrazolyl,benzothiofuranyl, benzothiazolyl, quinazolinyl and indazolyl, eachoptionally substituted with one or more halogen, —CN, alkylthio,alkylsulfinyl, alkylsulfonyl, —NO₂, SO₂NH₂ or R₇ where R₇ is C₁₋₆ alkyl,C₂₋₆ alkenyl, C₁₋₆ alkyloxyalkyl, C₁₋₄ alkyloxy, C₁₋₅ alkylamino, orC₁₋₆ alkylthioalkyl each optionally substituted with OH, CN, —COO-loweralkyl, —CONH-lower alkyl, —CON(lower alkyl)₂, dialkylamino, phenyl orheterocyclyl as hereinabove described in this paragraph; and R₈ is NH₂.11. The compound according to claim 10 wherein: R₁ is ethyl, isopropyl,n-propyl, t-butyl, cyclopentyl, CF₃, ethoxy, CH₃OCH₂—, 2- or3-tetrahydrofuranyl, 2-, 3-, or 4-pyridyl, 2-furanyl, or 2-thiazolyl; R₃is CN, CF₃, Cl, methyl, ethyl, SCH₃, cyclopropyl, vinyl or 2-furanyl; Lis —NHC(O)—, and R₄ is a phenyl or pyridyl each optionally substitutedwith one to three halogen, —CN, alkylthio, alkylsulfinyl, alkylsulfonylor R₇ where R₇ is C₁₋₆ alkyl, C₂₋₆ alkenyl, C₁₋₆ alkyloxyC₁₋₆ alkyl,C₁₋₄ alkyloxy, C₁₋₅ alkylamino each optionally substituted with halogen,OH, CN, COO-lower alkyl, —CONH-lower alkyl, —CON(lower alkyl)₂,dialkylamino, phenyl, morpholinyl or pyridyl.
 12. The accordingaccording to claim 11 wherein: R₁ is isopropyl, CF₃, 3-pyridyl or4-pyridyl; R₂ is H; R₃ is CN, CF₃, Cl, methyl, SCH₃ or ethyl; and R₄ isa phenyl or pyridyl each optionally substituted with one to three groupsselected from halogen, —CN, alkylthio, alkylsulfinyl, alkylsulfonyl orR₇ where R₇ is C₁₋₆ alkyl, C₁₋₄ alkyloxy, C₁₋₅ alkylamino eachoptionally substituted with OH, CN, COO-lower alkyl, —CONH-lower alkyl,—CON(lower alkyl)₂, dialkylamino, phenyl, morpholinyl or pyridyl.
 13. Acompound, wherein the compound is:

or the pharmaceutically acceptable derivatives thereof.
 14. Apharmaceutical composition comprising a therapeutically effective amountof a compound according to claim
 9. 15. A method of treating acardiovascular disease, said method comprising administering to apatient in need thereof a therapeutically effective amount of a compoundof according to claim
 9. 16. The method according to claim 15 whereinthe cardiovascular disease is involves endothelial dysfunction.
 17. Themethod according to claims 16 wherein the cardiovascular disease ischosen from type 1 and type 2 diabetes, insulin resistance syndrome,hypertension, atherosclerosis, coronary artery disease, angina,ischemia, ischemic stroke, Raynaud's disease and renal disease.